• Journal of Microbiology and Biotechnology
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• v.12 no.4
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• pp.706-709
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• 2002

Bacterial bioluminescence has been known to be a highly valuable reporter system for its potential application as an effective and simple environmental monitoring method for toxic compounds. In this short report, we constructed a streptomycetes-Escherichia coli shuttle vector-containing bioluminescence system and evaluated its potential application for toxic compounds monitoring. The luxAB biolurninescence genes from Vibrio harveyi were cloned into a streptornycetes-E. coli shuttle vector (named pESK004) and functionally expressed in Streptomyces lividans. The recombinant S. lividans containing pESK004 exhibited an optimal biolurninescence at the optical density ($OD_{600\;nm}$) of 0.4-0.5 and aldehyde concentration of 0.005%. When the recombinant bioluminescence streptomycetes was exposed to a toxic compound such as heavy metals, chlorinated phenols, or pesticides, the bioluminescence was decreased proportionally to the concentration of toxic compound in the assay mixture. The $EC_{50}$ (effective concentration to decrease 50% of the bioluminescence prior to exposure) values in the recombinant biolurninescence streptomycetes for mercury, 2,4-dichlorophenol, and malathion were measured at 2.2 ppm, 144.0 ppm, and 82.4 ppm, respectively. The degree of sensitivity and specificity pattern toward these toxic compounds characterized in this recombinant bioluminescence streptomycetes were unique when compared with previously reported bacterial bioluminescence systems, and this revealed that a recombinant bioluminescence streptomycetes might provide an alternative or complementary system for potential environmental monitoring.

• Korean journal of food and cookery science
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• v.16 no.2
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• pp.195-201
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• 2000

An investigation was conducted to evaluate the hygienic status of university foodservice operation by using conventional swabbing technique plus standard plate count and ATP bioluminescence assay. The results of the study were as follows: 1) For all kitchen boards, knives, feeding trays, and dish towels tested, there was an overall agreement at 84.7% level between the results obtained using ATP bioluminescence and plate count when using a pass/fail cut-off of 3$\times$ control values for ATP assay and 40 CFU(colony forming unit)/㎠ for plate count. 2) The agrement rate between ATP assay and standard plate count was 87.5% for the samples before use, 29.2% for those during use, and 42.7% for those after cleaning and sanitizing. 3) The plate counts of three university foodservice operations for kitchen board, kitchen knife, feeding tray and dish towel were within the acceptable limits when tested before using. However, none of them were within the acceptable limits when tested during using and after cleaning and sanitizing. 4) Above results suggested that an immediate action needs to be taken to reduce the potential danger of cross-contamination and also effective sanitary control methods needs to be developed to improve the sanitary condition.

• Journal of Korean Society of Environmental Engineers
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• v.30 no.1
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• pp.79-84
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• 2008

This paper describes the application of bioluminescence stimulating agents on a genetically engineered microorganism, Pseudomonas putida mt-2 KG1206, to monitor toluene analogs using in groundwater samples from petroleum hydrocarbon contaminated sites. The maximum bioluminescent response with pure chemicals followed in the order: m-methyl benzyl alchohol > m-toluate > toluene > m-xylene > benzoate > p-xylene > o-xylene. Generally, the bioluminescence production of strain mixed with groundwater samples was dependent on the contaminated total inducer concentrations. However, few samples showed opposite results, where these phenomena may be caused by the complexicity of environmental samples. Two chemicals, SL(sodium lactate) and KNO$_3$, were tested to determine a better bioluminescence stimulant. Both chemicals stimulate the bioluminescence activity of strain KG1206, however, a slightly high bioluminescence was observed with nitrogen chemical. This selected stimulant was then tested on samples collected from contaminated groundwater samples. The bioluminescence activity of all samples mixed with the strain was stimulated with KNO$_3$ amendment. This suggests that the low bioluminescence activity exhibited by the environmental groundwater samples can be stimulated by amending the culture with a proper agent, such as nitrogen compound. These findings would be useful, especially, when strain was used to monitor the groundwater samples contaminated with low inducer contaminants. Overall, the results of this study found the ability of bioluminescence producing bacteria to biosensor a specific group of environmental contaminants, and suggest the potential for more efficient preliminary application of this engineered strain in a field-ready bioassay.

• Journal of Microbiology and Biotechnology
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• v.16 no.11
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• pp.1706-1712
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• 2006

Because of the need for new antimicrobial substances with novel mechanisms of action, we report here the use of an Acinetobacter reporter system for high-throughput screening of active antimicrobial agents. The bioreporter Acinetobacter strain DF4/PUTK2 carrying luciferase genes luxCDABE was chosen because of its ecological importance and it is widespread in nature. This bioreporter is genetically engineered to emit light constitutively that can be measured in real time by luminometry. Hence, this reporter system was employed to determine the bacteriostatic actions of spent-culture supernatants derived from twelve bacterial isolates. Out of the results, the strongest bioluminescence inhibitory effect of the supernatants was recorded with Bacillus cereus strain BAC (S5). Subsequently, ethyl acetate extracts of extracellular products of strain BAC (S5) were separated by a thin-layer chromatography (TLC). Based on the bioluminescence inhibitory assay, three fractions were found to have antimicrobial activity. One fraction (C) having the strongest antimicrobial activity was further purified using TLC and characterized by IR, $^1H$ NMR, mass spectrometry, SDS-PAGE, and amino acid composition analysis. The results predicted the presence of 2-pyrrolidone-S-carboxylic acid (PCA) and the octadeconic-acid-like fatty acid. Fraction C also demonstrated a broad inhibitory activity on several Gram-negative and Gram-positive bacteria. In conclusion, the Acinetobacter reporter system shows great potential to be a reliable, sensitive, and real-time indicator of the bacteriostatic actions of the antimicrobial agents.

• Fisheries and Aquatic Sciences
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• v.15 no.3
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• pp.241-250
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• 2012

Polycyclic aromatic hydrocarbons (PAHs), butyltins (BTs), nonylphenol (NP), and fecal sterols concentrations in sediments were investigated from Gunsan coast of Korea to evaluate organic pollution from anthropogenic activities. Sediment toxicity was also examined by bacterial bioluminescence toxicity test (Vibrio fischeri). The concentrations of 16 PAHs in sediments ranged from 67.9 to 425 ng/g dry wt; BTs ranged from 2.79 to 14.1 ng Sn/g dry wt; NP ranged from 20.7 to 2171 ng/g dry wt; and coprostanol, a fecal sterol, ranged from 7.60 to 245 ng/g dry wt. Effective concentration 50% ($EC_{50}$) of sediments ranged from 0.38 to 23.8 mg/mL. Most of the chemicals were present at levels lower than or comparable to the previously reported values from Korea. However, NP levels in the present study were in the high range of levels reported from the Korean coast, and 40% of the measured samples exceeded screening and ecotoxicological values of NP suggested by the Netherlands and Canada. This suggests that an ongoing source of NP is a serious concern in the Gunsan coast. High levels of contaminants were found in the proximity of potential sources, such as the outfall of a wastewater treatment plant for NP, an anthracite-fired power plant for PAHs, and ports for BTs, fecal sterols, and sediment toxicity. This indicates that Gunsan coast has various potential sources of marine sediment contaminants.

• Toxicological Research
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• v.29 no.1
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• pp.1-6
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• 2013

The process of drug discovery and development requires substantial resources and time. The drug industry has tried to reduce costs by conducting appropriate animal studies together with molecular biological and genetic analyses. Basic science research has been limited to in vitro studies of cellular processes and ex vivo tissue examination using suitable animal models of disease. However, in the past two decades new technologies have been developed that permit the imaging of live animals using radiotracer emission, X-rays, magnetic resonance signals, fluorescence, and bioluminescence. The main objective of this review is to provide an overview of small animal molecular imaging, with a focus on nuclear imaging (single photon emission computed tomography and positron emission tomography). These technologies permit visualization of toxicodynamics as well as toxicity to specific organs by directly monitoring drug accumulation and assessing physiological and/or molecular alterations. Nuclear imaging technology has great potential for improving the efficiency of the drug development process.

• Journal of Marine Life Science
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• v.7 no.2
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• pp.74-85
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• 2022

Organisms constituting a large proportion of marine ecosystems, ranging from bacteria to fish, exhibit fluorescence and bioluminescence. A variety of marine organisms utilize these biochemically generated light sources for feeding, reproduction, communication, and defense. Since the discovery of green fluorescent protein and the luciferin-luciferase system more than a century ago, numerous studies have been conducted to characterize their function and regulatory mechanism. The unique properties of fluorescent and bioluminescent proteins offer great potential for their use in a broad range of applications. This short review briefly describes the functions and characteristics of fluorescent and bioluminescent proteins, in addition to summarizing the recent status of their applications.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2000.04a
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• pp.108-113
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• 2000

Freshwater borne bacteria transformed with luxAB-containing plasmid were optimized for the toxicity tests of various organic carbons and heavy metals. The EC$\sub$50/ values obtained from tests using the most sensitive bacterium to toxicants, YH9-RC, revealed to be much less than those from the Microtox$\^$/. In addition, some physiological characteristics of this bacterium under the toxic stress conditions such as potential bioluminescence, specific growth rate, and intracellular ATP contents, reproducibly and reliably correlated to the toxicity of the chemicals exposed. The higher concentrations of COD in wastewater samples, the lower EC$\sub$50/ values, therefore the developed toxicity test was found to be easily applicable to the toxicity test for wastewater samples and effluents. The conditions for constructing 384-multiwell plate containing freeze-dried bacterium were also optimized through the addition of 0.16 M trehalose before freeze-drying. Consequently, the advanced test system featuring a continuous measurement of the toxicity, an automated real-time monitoring of its results, and an alerting function was designed and constructed in combination with the microbiological, mechanical, and electronic compartment.

• Fisheries and Aquatic Sciences
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• v.13 no.3
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• pp.244-253
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• 2010

The quality of marine sediments from the industrialized coastal areas of Korea (Ulsan Bay, Masan Bay, and artificial Lake Shihwa) was investigated using a bacterial bioluminescence toxicity test. Sediment toxicity results were compared with the levels of chemical contamination (trace metals, organic wastewater markers, acid volatile sulfides, total organic carbon). Effective concentration 50% (EC50) of sediments ranged from 0.014 to 1.126 mg/mL, which is comparable to or lower than values in contaminated lakes, rivers, and marine sediments of other countries. Sediment reference index (SRI) ranged from 13 to 1044, based on the EC50 of the negative control sample. Mean average SRI values in Masan Bay and Lake Shihwa were approximately 8 and 9 times as high as that in Ulsan Bay, indicating higher sediment toxicity and greater contamination in the two former regions. Sediment toxicity were strongly associated with the concentrations of some chemicals, suggesting that this test may be useful for determining potential chemical contamination in sediments.

• Journal of Microbiology and Biotechnology
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• v.33 no.3
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• pp.329-338
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• 2023

Enterohemorrhagic Escherichia coli (EHEC) is a foodborne pathogen that produces attaching and effacing lesions on the large intestine and causes hemorrhagic colitis. It is primarily transmitted through the consumption of contaminated meat or fresh produce. Similar to other bacterial pathogens, antibiotic resistance is of concern for EHEC. Furthermore, since the production of Shiga toxin by this pathogen is enhanced after antibiotic treatment, alternative agents that control EHEC are necessary. This study aimed to discover alternative treatments that target virulence factors and reduce EHEC toxicity. The locus of enterocyte effacement (LEE) is essential for EHEC attachment to host cells and virulence, and most of the LEE genes are positively regulated by the transcriptional regulator, Ler. GrlA protein, a transcriptional activator of ler, is thus a potential target for virulence inhibitors of EHEC. To identify the GrlA inhibitors, an in vivo high-throughput screening (HTS) system consisting of a GrlA-expressing plasmid and a reporter plasmid was constructed. Since the reporter luminescence gene was fused to the ler promoter, the bioluminescence would decrease if inhibitors affected the GrlA. By screening 8,201 compounds from the Korea Chemical Bank, we identified a novel GrlA inhibitor named Grlactin [3-[(2,4-dichlorophenoxy)methyl]-4-(3-methylbut-2-en-1-yl)-4,5-dihydro-1,2,4-oxadiazol-5-one], which suppresses the expression of LEE genes. Grlactin significantly diminished the adhesion of EHEC strain EDL933 to human epithelial cells without inhibiting bacterial growth. These findings suggest that the developed screening system was effective at identifying GrlA inhibitors, and Grlactin has potential for use as a novel anti-adhesion agent for EHEC while reducing the incidence of resistance.